Expansion of rat 6-thioguanine-resistant T-lymphocyte clones by stimulation with ionomycin and a phorbol ester.
Previous molecular analyses of the mutations produced in the rat lymphocyte hprt assay were hindered by difficulties encountered in growing mutant lymphocytes from 6-thioguanine-resistant clones. In this study, we evaluated the ability of the calcium ionophore, ionomycin, and the tumor promotor, phorbol 12-myristate 13-acetate, to stimulate clone expansion. A medium containing these two agents, along with mitogen-free conditioned medium, was found to expand 64% of 276 mutant clones to at least 5 x 10(5) cells in nine days of culture. Some clones were expanded to more than 4 x 10(6) cells. The procedure appears suitable for propagating rat lymphocyte clones for mutation analysis.
Chen
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T
Division of Genetic and Reproductive Toxicology, National Center for Toxicological Research, Jefferson, Arkansas 72079, USA. TChen@NCTR.FDA.GOV
Aidoo
A
A
Casciano
D A
DA
Heflich
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RH
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells; D017077:Culture Media, Conditioned; D015759:Ionomycin / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects; D051381:Rats; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013866:Thioguanine / Q000494:pharmacology*
D017077:Culture Media, Conditioned; D015759:Ionomycin; D013866:Thioguanine; D013755:Tetradecanoylphorbol Acetate